Genome-wide localization of pre-RC sites and identification of replication origins in fission yeast

Makoto Hayashi, Yuki Katou, Takehiko Itoh, Mitsutoshi Tazumi, Yoshiki Yamada, Tatsuro Takahashi, Takuro Nakagawa, Katsuhiko Shirahige, Hisao Masukata

Research output: Contribution to journalArticle

139 Citations (Scopus)

Abstract

DNA replication of eukaryotic chromosomes initiates at a number of discrete loci, called replication origins. Distribution and regulation of origins are important for complete duplication of the genome. Here, we determined locations of Orc1 and Mcm6, components of pre-replicative complex (pre-RC), on the whole genome of Schizosaccharomyces pombe using a high-resolution tiling array. Pre-RC sites were identified in 460 intergenic regions, where Orc1 and Mcm6 colocalized. By mapping of 5-bromo-2′-deoxyuridine (BrdU)-incorporated DNA in the presence of hydroxyurea (HU), 307 pre-RC sites were identified as early-firing origins. In contrast, 153 pre-RC sites without BrdU incorporation were considered to be late and/or inefficient origins. Inactivation of replication checkpoint by Cds1 deletion resulted in BrdU incorporation with HU specifically at the late origins. Early and late origins tend to distribute separately in large chromosome regions. Interestingly, pericentromeric heterochromatin and the silent mating-type locus replicated in the presence of HU, whereas the inner centromere or subtelomeric heterochromatin did not. Notably, MCM did not bind to inner centromeres where origin recognition complex was located. Thus, replication is differentially regulated in chromosome domains.

Original languageEnglish
Pages (from-to)1327-1339
Number of pages13
JournalEMBO Journal
Volume26
Issue number5
DOIs
Publication statusPublished - Mar 7 2007

Fingerprint

Replication Origin
Hydroxyurea
Schizosaccharomyces
Chromosomes
Yeast
Deoxyuridine
Heterochromatin
Centromere
Genes
Genome
Origin Recognition Complex
Intergenic DNA
DNA
Multicarrier modulation
Bromodeoxyuridine
DNA Replication

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Genome-wide localization of pre-RC sites and identification of replication origins in fission yeast. / Hayashi, Makoto; Katou, Yuki; Itoh, Takehiko; Tazumi, Mitsutoshi; Yamada, Yoshiki; Takahashi, Tatsuro; Nakagawa, Takuro; Shirahige, Katsuhiko; Masukata, Hisao.

In: EMBO Journal, Vol. 26, No. 5, 07.03.2007, p. 1327-1339.

Research output: Contribution to journalArticle

Hayashi, M, Katou, Y, Itoh, T, Tazumi, M, Yamada, Y, Takahashi, T, Nakagawa, T, Shirahige, K & Masukata, H 2007, 'Genome-wide localization of pre-RC sites and identification of replication origins in fission yeast', EMBO Journal, vol. 26, no. 5, pp. 1327-1339. https://doi.org/10.1038/sj.emboj.7601585
Hayashi, Makoto ; Katou, Yuki ; Itoh, Takehiko ; Tazumi, Mitsutoshi ; Yamada, Yoshiki ; Takahashi, Tatsuro ; Nakagawa, Takuro ; Shirahige, Katsuhiko ; Masukata, Hisao. / Genome-wide localization of pre-RC sites and identification of replication origins in fission yeast. In: EMBO Journal. 2007 ; Vol. 26, No. 5. pp. 1327-1339.
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